123 results about "Cell factory" patented technology

The invention discloses a novel biosynthesis path for producing 3 '-phosphoadenosine-5'-phosphosulfuric acid and application, and belongs to the technical field of bioengineering. According to the invention, through recombinant expression and purification of ribose kinase, ribose phosphate pyrophosphate kinase, ribose phosphate transferase, adenine thiotransferase, adenosine phosphate sulfate kinase and polyphosphate kinase 2, a six-enzyme cascade reaction taking ribose and adenosine as substrates in vitro is constructed, the molar conversion rate reaches 22.85%, and the escherichia coli and bacillus subtilis PAPS cell factory can be used for preparing the recombinant ribose phosphate pyrophosphate kinase, the ribose phosphate transferase, the adenine thiotransferase, the adenosine phosphate sulfate kinase and the polyphosphate kinase 2. The yield of PAPS in a 5-L fermentation tank reaches 7.60 g / L and 5.03 g / L respectively. The PAPS synthesis route provided by the invention provides an extensible, economical and efficient solution for the production of sulfated compounds.

The invention discloses a novel plasmid vector capable of increasing CHO foreign protein expression quantity and construction and application thereof, and belongs to the technical field of gene engineering. The plasmid vector pCHO6GS belongs to a brand-new plasmid system, and the expression quantity of a common monoclonal antibody and recombinant protein (such as H5HA trimer) difficult to express in CHO can be increased by at least more than one time, so that the expression quantity of foreign protein is increased beyond expectation through the novel plasmid vector; and the method also plays an extremely important role in reducing cost and improving efficiency of various protein products taking CHO as a cell factory.

The present disclosure relates to a genetically modified host cell having improved production of sulfurol and / or sulfurol precursors, wherein the host cell expresses one or more heterologous thiaminase I, thiaminase II and / or phosphatase enzymes and / or one or more heterologous thiamine pathway enzymes converting thiamine, 5-(2-hydroxyethyl)-4-methyl-1,3-thiazole-2-carboxylic phosphate acid (cTHZ-P), 5-(2-hydroxyethyl)-4-methyl-1,3-thiazole-2-carboxylic acid (cTHZ) and / or THZ-P to sulfurol and / or 5-(2-hydroxyethyl)-4-methyl-1,3-thiazole-2-carboxylic acid (cTHZ), whereby the production of the sulfurol in the genetically modified host cell is improved compared to an unmodified parent host cell. Chemical methods for production of sulfurol are also provided.

The invention discloses a bacillus constitutive strong promoter and application thereof, and relates to the technical field of biology, and the nucleotide sequence of the promoter is SEQ ID NO: 2. The mRNA transcriptional level of the reporter gene is improved; the method is used for starting a target gene to efficiently express amylase in bacillus subtilis. The method is also used for starting a target gene to efficiently express alkaline protease in bacillus licheniformis. The novel constitutive promoter with enhanced transcriptional activity is developed by taking P43 as a starting template, and under the same host and culture conditions, the absolute level of the constitutive promoter for driving expression of a reporter gene or a target product is higher than that of the current P43 promoter, so that core power is provided for performance improvement of a bacillus cell factory.

The invention provides a cell strain for producing engineered viroid particles, a method thereof and the engineered viroid particles, the cell strain is named as SZCV-Con28 and preserved in Guangdong Microbial Culture Collection Center on October 15, 2025, the preservation number is GDMCC No: 67101, and the preservation number is GDMCC No: 67101. A plurality of corresponding cell factories for producing the engineered viroid particles are established by cell lines obtained by introducing different target gRNA elements into the cell strain, so that the corresponding engineered viroid particles can be continuously produced in batches. According to the invention, large-scale preparation can be realized through cell expanding culture, the capacity limitation is overcome, the production process flow is simplified, and the consumption of production raw materials is reduced.

The invention discloses curcuma wenyujin sesquiterpene synthase CwTPS8 as well as a coding gene and application thereof, and belongs to the technical field of plant genetic engineering, the amino acid sequence of the curcuma wenyujin sesquiterpene synthase CwTPS8 is shown as SEQ ID NO.1, the nucleotide sequence of the gene for coding the curcuma wenyujin sesquiterpene synthase CwTPS8 is shown as SEQ ID NO.2, and the nucleotide sequence of the gene for coding the curcuma wenyujin sesquiterpene synthase CwTPS8 is shown as SEQ ID NO.1. The curcuma wenyujin sesquiterpene synthase CwTPS8 can be used for preparing beta-caryophyllene, alpha-humulene and other curcuma wenyujin sesquiterpenoids by taking farnesyl pyrophosphate FPP as a substrate, and the technology can be used for subsequently producing the sesquiterpenoids through a microbial cell factory. Meanwhile, a foundation is laid for heterologous synthesis and further development and utilization of sesquiterpenoids such as beta-caryophyllene and alpha-humulene.

This invention aims to create a spatially isolated local reaction microenvironment for Rubisco by introducing a complete β-carboxysome microcompartment system. This design goal avoids oxygen inhibition, laying the foundation for ultimately achieving efficient and specific CO2 fixation. It provides a novel approach to overcome the fundamental problem of low efficiency of carbon-fixing enzymes in cytosols due to intense oxygen competition. By introducing the complete carboxysome system as a module, engineered yeast strains with high carboxylase activity and significantly improved ethanol production were obtained, providing a solid foundation for building next-generation yeast cell factories that produce high-yield ethanol using CO2 as a raw material.

This invention relates to the field of synthetic biology, and more particularly to a microbial cell factory for producing catechols. The invention uses *Pseudomonas putida* strains with the pcaHG, catA, and catA2 genes knocked out as substrate bacteria, and introduces exogenous AroY and / or kpdB genes, or increases the expression of endogenous genes VanAB and PobA. Through a strategy of cofactor regulation and key enzyme overexpression, lignin-derived monomers are efficiently converted into catechols, achieving a conversion rate of 98.5% and a maximum yield of 14.1 mM, a significant improvement compared to the previous conversion rate of 64%.

The invention relates to a high-titer influenza virus seed as well as a preparation method and application thereof, and the preparation method comprises the following steps: S1, obtaining a virus, inoculating the virus into a cell factory, and culturing to obtain a cell supernatant; and S2, carrying out microfiltration on the cell supernatant to remove cell debris in the cell supernatant, carrying out ultrafiltration concentration, and carrying out degerming filtration to obtain the high-titer influenza virus seed. According to the technical scheme, the cell factory is adopted for culturing the virus, the virus is inoculated to the ten-layer cell factory, after culturing, the cell supernatant in the cell factory is harvested, microfiltration is carried out, and the cell debris is removed, so that compared with a conventional method for removing the cell debris through centrifugation, microfiltration needs shorter time, the volume of the supernatant which can be treated at a time is larger, and the cell debris can be removed at a time. The sterile risk is lower; and concentrating the supernate, and sterilizing and filtering after concentration. The virus titer of the virus seed subjected to the preparation process can be increased by 5-10 times, and the impurity content is lower.

A simpler and lower-cost method and associated hardware are described for recovering biopharmaceutical products from cell factories in higher yields by a liquid nitrogen-based pre-treatment of cell factories followed by gentler disintegration of cellular factories by tunable, pressurizing and de-pressurizing cycle with gaseous nitrogen. The cell factories are heterologous expression systems and could be based on plant cells, mammalian cells, algal cells, yeast cells or even bacterial cells. The pre-treatment step with liquid nitrogen softens the tougher cell wall structures thus enabling cells to be disrupted relatively easily during the gaseous nitrogen-based pressurizing / depressurizing cycle. This invention can enhance the yield of biopharmaceutical products from expression systems and reduce the overall cycle-time and further down-stream processing and polishing cost of biopharmaceutical products.

The invention relates to polymerized glutamate sugar ester as well as a biological fermentation preparation method and application thereof, and belongs to the technical field of biological fermentation. According to the scheme, amino acid and functional oligosaccharide are taken as substrates, and efficient directional assembly of high-molecular polymerized glutamate sugar ester is realized by designing an integrated cell factory biosynthesis technology. The invention provides a brand-new polyglutamic acid sugar ester compound and a biological fermentation method of the polyglutamic acid sugar ester, the biological fermentation method is suitable for industrial continuous production, and the prepared polyglutamic acid sugar ester is narrower in molecular weight distribution and better in product uniformity.

The application discloses a kind of biological immobilized enzyme based on yeast surface display technology and the application of synthesis 2-phenylethanol, using yeast surface display technology, for the first time with Saccharomyces cerevisiae surface protein a-lectin as anchor protein, the key enzyme (transaminase, decarboxylase, ethanol dehydrogenase) involved in 2-PE producing Ehrlich pathway in Saccharomyces cerevisiae is separately or jointly carried out biological fixation, enhance substrate channel effect and proximity effect, in vitro one-step catalysis L-phenylalanine synthesis 2-PE, provide yield.The method for fixing multiple enzymes of Ehrlich pathway in the application can be used to reconfigure the biosynthesis pathway of other natural products in vitro, to provide feasible scheme for the rational design of cell factory.

The application belongs to the technical field of microbial sewage treatment, and particularly relates to an iron-carbon quantum dot / electrically active non-oxygen-producing photosynthetic bacteria / carrier hybrid cell factory and application thereof in sewage resourceization or carbon emission reduction; the iron-carbon quantum dot in the hybrid cell factory provided by the application promotes the non-oxygen-producing purple photosynthetic bacteria to utilize more light energy, and generates photo-generated electrons as an extracellular efficient electron donor source, and the photosensitive active substances in the extracellular polymeric substance of the non-oxygen-producing purple photosynthetic bacteria can promote the formation of a continuous cycle of trivalent iron and divalent iron, realize mutual cooperation, promote sewage resourceization and carbon emission reduction, and synthesize high-value-added products; and in addition to facilitating the recovery of biomass, the transparent conductive glass electrode as the carrier also ensures that the non-oxygen-producing purple photosynthetic bacteria stably and efficiently utilize light energy; therefore, the hybrid cell factory provided by the application is a high-efficiency microbial sewage treatment technology, and solves the technical problem of lack of high-efficiency microbial sewage treatment technology in the prior art.

The present disclosure relates to genetically modified host cells producing prostaglandins, to genetic constructs for expression of such mutants; to cultures of the genetically modified host cells and its use to produce the said products; to fermentation liquids comprising the said products resulting from such production; to compositions comprising the fermentation liquid; to pharmaceutical preparations made from such compositions and to the uses of such compositions and preparations.

The invention discloses a double-fucosyl transferase complex and application thereof in synthesis of difucosyl lactose, and belongs to the technical field of biology. In order to solve the problem of high residual quantity of a by-product 2 '-FL in the existing difucosyl lactose (DFL) cell factory, the invention constructs a novel difucosyl transferase complex, random mutation and screening are performed on a key element connecting peptide in the complex, and the catalytic effect of an enzyme is optimized. Through the strategy, the constructed recombinant strain can control the residual quantity of a byproduct 2 '-FL while efficiently synthesizing the DFL, so that the subsequent separation and purification cost is remarkably reduced, the production efficiency is improved, and important data support is provided for industrial large-scale production of the DFL.

The utility model relates to cell culture technical field discloses a kind of cell factory device, including plug body and the cover disc and culture tray of stacked arrangement, cover disc is located the top of culture tray, culture tray includes culture bottom plate, culture side plate and communicating part, culture side plate is arranged along the edge of culture bottom plate, and one end of culture side plate is connected with culture bottom plate and is enclosed to form culture cavity, and the other end of culture side plate is connected with culture tray, and culture tray seals culture cavity, culture bottom plate is equipped with through-hole, and communicating part is located in culture cavity, and one end of communicating part is connected in through-hole, and the other end of communicating part is formed with flow port between culture tray, cover disc is equipped with liquid injection hole, and the position of liquid injection hole is correspondingly set with the position of communicating part, and liquid injection hole is communicated with culture cavity by flow port, and plug body is sealedly connected with one end of communicating part away from cover disc.The cell factory device of the utility model improves the control of flatness of cell factory device, and ensures that cell growth is evenly distributed.

The invention discloses a construction method and application of a kluyveromyces marxianus mutant, and belongs to the field of synthetic biology and molecular biology. According to the method, single-stranded DNA binding protein and cytosine deaminase pmCDA1 are fused, a random base editing system is constructed, and whole-genome efficient C-T / G-A mutation is realized in kluyveromyces marxianus by taking MCM2-7 as a verification target. The system does not need a specific target or an exogenous DNA template, the mutation efficiency is high, the distribution is wide, the mutation rate is improved by 303.1 times compared with that of a control group through optimal combination, and the application potential of kluyveromyces marxianus in industrial production is expanded. The method is combined with an efficient screening system, mutant strains with the intracellular protein expression quantity remarkably improved are screened out, the constructed mutation system is not only suitable for kluyveromyces marxianus cell factory transformation, but also can be popularized to genome evolution of other yeasts or eukaryotic hosts, and a new strategy is provided for industrial microorganism breeding and protein engineering.

The application relates to a polymeric glutamic acid sugar ester and a biological fermentation preparation method and application thereof, and belongs to the technical field of biological fermentation. According to the application, amino acids and functional oligosaccharides are used as substrates, and a 'one-body cell factory' biosynthesis technology is designed to realize efficient directional assembly of high-molecular polymeric glutamic acid sugar esters. The application provides a brand-new polymeric glutamic acid sugar ester compound, and provides a biological fermentation method of the polymeric glutamic acid sugar ester, which is suitable for industrialized continuous production, and the prepared polymeric glutamic acid sugar ester has a narrower molecular weight distribution and better product uniformity.

The embodiment of the invention provides a cell factory reactor and a use method, and relates to the technical field of cell culture, the cell factory reactor comprises: a cell factory main body having a plurality of culture layers and a first channel respectively communicated with the plurality of culture layers, each culture layer having an upper surface and a lower surface, both the upper surface and the lower surface being subjected to TC treatment; the sensing assembly is used for acquiring culture environment parameters in the cell factory main body; the process gas supply assembly is connected with the first channel; the pipeline system comprises a liquid input assembly and a liquid output assembly, the liquid input assembly is used for conveying materials required by cell culture to the first channel, and the liquid output assembly is used for transferring liquid or cell suspension in the cell factory main body to the outside of the cell factory main body; the controller comprises a swinging mechanism, and the swinging mechanism is used for driving the cell factory main body to swing. According to the embodiment of the invention, the scale of cell amplification is increased, and the problem of insufficient amplification scale of a traditional cell factory is effectively solved.

The invention relates to the technical field of microbial fermentation and genetic engineering, and particularly discloses a yeast engineering bacterium for efficiently producing plastic depolymerases and application of the yeast engineering bacterium. The engineering yeast takes pichia pastoris as an expression host, a molecular chaperone is co-expressed, a vacuole sorting receptor gene is knocked out or knocked down, in addition, a yeast cell factory for efficiently expressing the plastic depolymerases is constructed by further combining regulation and control element optimization and target gene copy number regulation, and the engineering yeast has the advantages that the engineering yeast can be used for efficiently expressing the plastic depolymerases; the regulatory element comprises any one or a combination of more of a strong promoter, a signal peptide and a C-terminal tag. The engineering yeast provided by the invention can realize efficient accumulation and secretion of the plastic depolymerases, obviously improve the enzyme yield and reduce the production cost of unit enzyme activity, and provides a stable and efficient yeast expression system for industrial and low-cost production of the plastic depolymerases; the method has an important application prospect in the fields of biodegradation and resource utilization of waste plastics.

This invention discloses a method for designing optimal strains for product synthesis using an enzyme-constrained model combined with an evolutionary algorithm, comprising the following steps: Step S1, solving the enzyme-constrained model with the objective function of maximizing the specific growth rate of the strain to obtain the simulated metabolic flux of the wild-type strain; Step S2, performing dimensionality reduction analysis and labeling on genes directly related to enzyme synthesis within the model by solving a series of flux balance analysis problems with fixed biomass synthesis rates; Step S3, predicting the yield of single-target editing; Step S4, using a genetic algorithm to search for combined targets after dimensionality reduction, adjusting relevant parameters to obtain the optimal gene editing combination strategy; Step S5, repeating the experiment to verify the stability of the algorithm. This invention utilizes genetic algorithms and enzyme-constrained models to discover non-intuitive gene editing combination strategies, providing a new method for constructing efficient microbial cell factories for synthetic chemicals, and demonstrating the potential of applying heuristic algorithms to strain optimization design.

This invention relates to a Bacillus licheniformis strain with high tolerance to methanol and formaldehyde, belonging to the fields of synthetic biology and microbial engineering. This invention utilizes systematic mutagenesis and directed evolution to screen for a Bacillus licheniformis strain exhibiting excellent dual tolerance to methanol and formaldehyde. Bacillus licheniformis GW-MF, accession number CGMCC No. 38523. This strain can tolerate methanol concentrations of 80 g / L or even higher and formaldehyde concentrations of 0.02 g / L. This strain provides a key platform for constructing methanol-feeding cell factories, suitable for the green manufacturing process of methanol bioconversion to synthesize high-value-added products. It meets the development needs of a one-carbon bioeconomy under the "dual carbon" goal and has great application potential.

The application discloses a novel plasmid vector with improved CHO exogenous protein expression amount and construction and application thereof, and belongs to the technical field of genetic engineering. The plasmid vector pCHO6GS belongs to a novel plasmid system, and can at least improve the expression amount of general monoclonal antibodies and difficult-to-express recombinant proteins (such as H5HA trimer) in CHO by more than 1 times, that is, the unexpected improvement of the exogenous protein expression amount is realized by the novel plasmid vector, and the novel plasmid vector has an extremely important role in cost reduction and efficiency increase of various protein products with CHO as a cell factory.

This invention discloses a method for constructing a high-succinic acid-producing Pichia pastoris cell factory, using Pichia pastoris (… Komagataella phaffii Using GS115 as the chassis, a reductive TCA cycle enhancement strategy and an sdh5 knockout strategy were first constructed, respectively. It was found that a single strategy was insufficient to effectively achieve succinic acid accumulation. Further, the two strategies were combined to obtain the engineered strain SAOG, which achieved succinic acid accumulation. Based on this, the malate transporter SpMAE was introduced to enhance the transmembrane transport capacity of succinic acid, and histidine auxotrophic recovery was combined to improve the growth and fermentation performance of the strain, resulting in the engineered strain SH, whose succinic acid accumulation was 25.1% higher than that of SAOG. Finally, the engineered strain SH achieved a succinic acid yield of 10.47 g / L in shake-flask fermentation and 112.6 g / L after 120 h of continuous fed-batch fermentation in a 5 L fermenter. This invention achieves efficient succinic acid synthesis in the Pichia pastoris chassis through the synergistic regulation of reductive TCA pathway enhancement, succinic acid oxidation blocking, enhanced transmembrane transport, and auxotrophic recovery, exhibiting stable fermentation performance, high yield, and good industrial application potential.

The invention provides a biosynthesis method of 6-hydroxy-L-tryptophan, on the basis that a novel tryptophan 6-site hydroxylase protein family enzyme is obtained through screening, a cell factory is constructed for biotransformation, and green synthesis of 6-hydroxy-L-tryptophan is achieved. The technical problems of limited types of biocatalysts, insufficient chemical synthesis region selectivity and the like in the prior art are effectively solved. The amino acid sequence of the tryptophan 6-site hydroxylase is shown as SEQ ID NO: 1. The invention provides a protein sequence for catalyzing a new family of tryptophan 6-site hydroxylase, and 6-hydroxy-L-tryptophan can be directly and efficiently synthesized through a cell factory. The method has the advantages of mild reaction conditions, environmental friendliness, high regioselectivity, good product purity, flexible process route and the like, and a brand new technical scheme is provided for industrial production of 6-hydroxy-L-tryptophan.

The utility model discloses a microscopic observation system for a cell factory. The microscopic observation system comprises a main body frame, a front-back translation mechanism, a lifting mechanism, a focusing movement mechanism and a microscope observation module, a front-back translation mechanism and a lifting mechanism are arranged in the main frame from bottom to top, the lifting mechanism penetrates through the top surface of the main frame, a supporting platform is arranged at the top of the lifting mechanism, and the cell factory is arranged on the supporting platform; focusing moving mechanisms are arranged on the top surface of the main body frame and located on the left side and the right side of the supporting platform respectively, and microscope observation modules are installed above the focusing moving mechanisms; the system further comprises a display control module. And the display control module is electrically connected with the lifting mechanism, the front-back translation mechanism, the focusing movement mechanism and the microscope observation module. According to the utility model, automatic up-and-down and front-and-back movement of the cell factory and left-and-right movement of the microscope observation module are realized for observation operation, culture, growth cultivation and lesion microscope observation of the cell factory are realized, the automation degree is high, the overall integration degree is high, and the occupied volume is small.

The application discloses high-temperature-resistant engineered methanotrophs with high glutamic acid yield, a construction method and application thereof, and belongs to the field of industrial biotechnology. Methylotuvimicrobium buryatense 5GB1 as the host strain, transcriptional inhibition of the SOS response system by LexA protein is removed by knocking out a lexA gene in the genome of the host strain, expression of DNA damage repair related genes in the cell is activated, and high-temperature-resistant engineered methanotrophs MBΔ lexA that can stably grow in a high-temperature environment of 40-45 DEG C are obtained. The strain can still maintain normal growth and metabolism with methane as the sole carbon source under the condition of 40-45 DEG C, and the glutamic acid synthesis capacity of the strain is significantly better than that of the original strain at normal temperature. The high-temperature-resistant engineered methanotrophs MBΔ lexA provide important strain resources for the production of amino acid products by methanotroph cell factories in a high-temperature industrial scene, and are expected to improve the economy and sustainability of methanotrophic bioconversion.

The invention discloses a glycosylation-free human enterokinase mutant as well as recombinant expression and application thereof, belongs to the field of genetic engineering and protein technical engineering, and solves the problems of disordered glycosylation modification, low enzymatic activity and low protein expression quantity during recombinant expression of enterokinase in an enterokinase microbiological cell factory. The method comprises the following steps: firstly, carrying out single-site mutation, double-site mutation, three-site mutation and four-site mutation on the basis of a human enterokinase wild type, and constructing a pichia pastoris cell factory to obtain a series of human enterokinase mutants without glycosylation modification. Through calculation design and rational transformation, partial or non-glycosylation is subjected to deep fixed-point transformation, and pichia pastoris cell factory expression is constructed, so that the expression quantity and the total enzyme activity of the mutant are greatly improved. The highest enzyme activity of the glycosylation-free enterokinase mutant generated by fed-batch fermentation in a pichia pastoris cell factory reaches 7.8 * 10 < 4 > U / L, and the enterokinase mutant has the capacity of large-scale industrial fermentation generation and application.

The invention discloses a method for enhancing the synthesis capacity of a yeast double-metabolic-pathway pigment product and application, and belongs to the field of synthetic biology and metabolic engineering. The method comprises the following steps: constructing a saccharomyces cerevisiae engineering strain capable of simultaneously synthesizing violacein and lycopene; constructing a mutant library of the global transcription factor SPT15; transforming the mutant library into the engineering strain, and screening the mutant strain with increased yield based on bacterial colony color visualization. According to the invention, a single global transcription factor SPT15 is modified, so that the cell metabolism is regulated and controlled on the whole, the synergistic interaction of a diheterologous synthesis pathway is realized, and the tedious multi-gene rational modification is avoided. The method is easy and convenient to operate and short in period, and a new strategy is provided for efficient construction of yeast cell factories for high-yield pigment and other double-way products.

The invention discloses a genetically engineered bacterium for biosynthesis of inositol and application of the genetically engineered bacterium for biosynthesis of inositol, and the genetically engineered bacterium for biosynthesis of inositol is obtained by co-expressing a sweet orange inositol-3-phosphate synthase gene Csino3 and an escherichia coli inositol-1-monophosphate gene suhB in recombinant escherichia coli and then knocking out a glucose phosphate isomerase gene pgi. According to the technical scheme, the yield of inositol produced by the strain reaches 2.04 g / L and is increased by about 6.6 times compared with that before optimization, the inositol yield is greatly increased, the metabolic flux of glycolysis to glucose-6-phosphoric acid is effectively blocked, carbon metabolic flux is forced to be redirected to a pentose phosphate pathway, glucose-6-phosphoric acid is enriched, and a raw material is provided for inositol production. By blocking downstream branches of a glycometabolism pathway, enrichment of glucose-6-phosphate in cells is realized, so that sufficient precursor substances are provided for synthesis of inositol, and finally the yield of inositol is increased. The technical scheme is expected to be applied to construction of microbial cell factories, so that large-scale and low-cost production of inositol is realized, and theoretical support is provided for development of related industries.